RÉSUMÉ
Busulfan is an antineoplastic, which is always accompanied with the abnormal of spermatogonia self-renewal and differentiation. It has been demonstrated that the omega-3 polyunsaturated fatty acids (PUFAs) benefits mature spermatozoa. However, whether omega-3 can protect endogenous spermatogonia and the detailed mechanisms are still unclear. Evaluate of spermatogenesis function (in vivo) were examined by histopathological analysis, immunofluorescence staining, and western blotting. The levels of lipid metabolites in testicular tissue were determined via liquid chromatography. We investigated the effect of lipid metabolites on Sertoli cells provided paracrine factors to regulate spermatogonia proliferation and differentiation using co-culture system. In our study, we showed that omega-3 PUFAs significantly improved the process of sperm production and elevated the quantity of both undifferentiated Lin28+ spermatogonia and differentiated c-kit+ spermatogonia in a mouse model where spermatogenic function was disrupted by busulfan. Mass spectrometry revealed an increase in the levels of several omega-3 metabolites in the testes of mice fed with omega-3 PUFAs. The eicosapentaenoic acid metabolite 12-hydroxyeicosapentaenoic acid (12-HEPE) up-regulated bone morphogenic protein 4 (BMP4) expression through GPR120-ERK1/2 pathway activation in Sertoli cells and restored spermatogonia proliferation and differentiation. Our study provides evidence that omega-3 PUFAs metabolite 12-HEPE effectively protects spermatogonia and reveals that GPR120 might be a tractable pharmacological target for fertility in men received chemotherapy or severe spermatogenesis dysfunction.
Sujet(s)
Busulfan , Sperme , Humains , Mâle , Souris , Animaux , Busulfan/pharmacologie , Busulfan/métabolisme , Spermatogenèse/physiologie , Spermatogonies , Spermatozoïdes , Testicule/métabolismeRÉSUMÉ
PURPOSE: Non-obstructive azoospermia (NOA) is an essential cause of male infertility for which treatment options are limited. The pathogenic mechanism of NOA, especially idiopathic NOA, remains unclear. Gene variations are associated with the occurrence of NOA. Our study was performed to investigate the genetic causes of NOA. METHODS: Whole exome sequencing (WES) was performed in two probands diagnosed with NOA from a Chinese family. Sanger sequencing was applied to verify the pathogenic variants. A minigene assay was carried out to identify the effect of the splicing variants. RESULTS: We detected a novel homozygous variant (c.2681-3 T > A) in the HFM1 gene in the two siblings diagnosed with NOA, and their parents carried heterozygous mutations in the same gene. The results of the minigene assay revealed this splicing variant results in exon25 of HFM1 being skipped, leading to a protein truncation (p.Trp894Cysfs*44). CONCLUSION: Our results showed that a deleterious splicing variant in HFM1 was related to NOA in these two patients. This novel variant of HFM1 may serve as a potential genetic biomarker for NOA patients.
Sujet(s)
Azoospermie , Infertilité masculine , Humains , Mâle , Azoospermie/anatomopathologie , Infertilité masculine/génétique , Mutation/génétique , Méiose/génétiqueRÉSUMÉ
OBJECTIVE: To study the correlation, consistency, and variations between two assays of DNA fragmentation index based on acridine orange (AO) staining via AI-based fluorescence microscopy(AI-DFI), and flow cytometry (FCM-DFI) across multiple centers. METHODS: We selected 421 male patients from Nanjing Drum Tower hospital ( Hospital G) (226 cases), Eastern Theatre General Hospital (Hospital J) (89 cases) and Jiangsu Province Hospital (Hospital S) (106 cases) . Semen samples from each patient were analyzed for routine semen parameters and for DFI using both AI fluorescence microscopy and flow cytometry. We studied the two methods' stability as well as the correlation, consistency, and variation between the two methods' results in various centers. RESULTS: The two replicate studies' results of AI-DFI and the three centers' FCM-DFI for linear regression analysis indicated strong stability (R2>0.9).Overall(Group A), the AI-DFI results demonstrated good correlation and consistency with the FCM-DFI results of three centers (r>0.85ï¼ICC>0.9).The semen specimens were categorized into two groups: normal specimen group (group B) and abnormal specimen group (group C) (including asthenozoospermia, oligospermia, and semen samples with high impurities).Group C's results showed a decline in correlation and consistency when compared to group A and group B, whereas group B's results showed a little rise in correlation and consistency when compared to group A. Although the consistency and correlation between the results of the two DFI testing methods in the three centers were good, there was still a significant difference between Groups A and C (P<0.05), and in Group B there was a significant difference between the two DFI testing methods only in Hospital G (pï¼0.02), with no significant difference in Hospitals J and S (P> 0.05). CONCLUSION: The two detection methods exhibit good stability and correlation. However, significant differences are observed in the DFI detection methods in samples with abnormal semen parameters and high complexity. The main reason for these significant differences may lie in the variations in detection principles. Each detection method has its own advantages, allowing clinical or research settings to choose between them based on laboratory conditions or specific requirements.
Sujet(s)
Infertilité masculine , Sperme , Humains , Mâle , Analyse du sperme/méthodes , Cytométrie en flux/méthodes , Fragmentation de l'ADN , Spermatozoïdes , Microscopie de fluorescence , Coloration et marquage , Intelligence artificielle , Infertilité masculine/diagnostic , Infertilité masculine/génétiqueRÉSUMÉ
Obtaining high-quality sperm is key to improving the success rate of assisted reproductive technology (ART). Although cytokines secreted by cumulus-oocyte complexes (COCs) bind to sperm surface receptors to improve sperm quality, the effects of adding mouse COCs to human tubal fluid (HTF) medium on sperm capacitation have not yet been explored. Eight-week-old ICR mouse COCs were added to HTF medium and crushed to obtain the post-modified HTF medium. Compared with using HTF medium, the fertilisation rate and number of sperm combined with the zona pellucida significantly increased after in vitro capacitation using the post-modified HTF medium (P < 0.01). Proteomic and Western blotting analyses showed that the level of SERPINA5 in sperm increased significantly following in vitro capacitation with the post-modified HTF medium (P < 0.05). Immunohistochemical staining analysis demonstrated that SERPINA5 protein was expressed in mouse cumulus cells. A SERPINA5 antibody was added in the post-modified HTF medium to block the effects of SERPINA5 after in vitro capacitation, which significantly decreased the fertilisation rate and the number of sperm combined with the zona pellucida (P < 0.05). Recombinant mouse SERPINA5 protein (1 ~ 2 µg/ml) was added to HTF medium and the fertilisation rate and the number of sperm combined with the zona pellucida significantly increased (P < 0.01). Moreover, recombinant human SERPINA5 protein (5 µg/ml) was added before human semen freezing. Compared with adding no SERPINA5 protein, the percentage of normal sperm morphology and the intact acrosome significantly increased (P < 0.05). Our study provides a reference method for optimising sperm quality in the process of in vitro capacitation.
Sujet(s)
Inhibiteur de la protéine C , Sperme , Animaux , Femelle , Fécondation , Humains , Mâle , Souris , Souris de lignée ICR , Ovocytes , Inhibiteur de la protéine C/métabolisme , Protéomique , Capacitation des spermatozoïdes , Interaction sperme-ovule , Spermatozoïdes/métabolisme , Zone pellucide , Glycoprotéines de la zone pellucideRÉSUMÉ
Objective: To analyze the correlation of the sperm DNA fragmentation index (DFI) level with semen parameters and pregnancy outcomes of artificial insemination of the husband (AIH) in the cycle of intrauterine insemination (IUI). METHODS: We collected the clinical data on 777 cases of IUI, including female clinical indicators, male semen parameters, sperm DFI and pregnancy outcomes. According to the DFI level, we divided the patients into three groups: DFI < 15%, 15% ≤ DFI < 30% and DFI ≥ 30%. RESULTS: The sperm DFI level was significantly elevated with the increased age of the males (P = 0.002) and closely related to the total number of motile sperm (P = 0.002) and total sperm motility (P = 0.000) before treatment, as well as to sperm concentration (P = 0.000), total sperm motility (P = 0.001) and total number of progressively motile sperm (P = 0.000) after density gradient centrifugation. The rate of clinical pregnancy was decreased in the DFI ≥ 30% group. There were no statistically significant differences between sperm DFI and the rates of clinical pregnancy and abortion. CONCLUSIONS: Male age significantly affects the sperm DFI level. Sperm DFI is closely related to sperm motility and total number of progressively motile sperm, but not to the rates of clinical pregnancy and abortion in patients undergoing IUI. IUI can be used as an effective method of assisted reproduction for male infertility./.
Sujet(s)
Issue de la grossesse , Sperme , Fragmentation de l'ADN , Femelle , Humains , Insémination artificielle avec conjoint , Mâle , Grossesse , Mobilité des spermatozoïdes , SpermatozoïdesRÉSUMÉ
An increasing number of men are fathering children at an older age than in the past. While advanced maternal age has long been recognized as a risk factor for adverse reproductive outcomes, the influence of paternal age on reproduction is incompletely comprehended. Herein, we found that miR-125a-5p was upregulated in the sperm of aging males and was related to inferior sperm DNA integrity as an adverse predictor. Moreover, we demonstrated that miR-125a-5p suppressed mitochondrial function and increased cellular DNA damage in GC2 cells. We also found that miR-125a-5p perturbed embryo development at specific morula/blastocyst stages. Mechanistically, we confirmed that miR-125a-5p disturbed the mitochondrial function by targeting Rbm38 and activating the p53 damage response pathway, and induced a developmental delay in a p21-dependent manner. Our study revealed an important role of miR-125a-5p in sperm function and early embryo development of aging males, and provided a fresh view to comprehend the aging process in sperm.
Sujet(s)
Altération de l'ADN/génétique , Développement embryonnaire/génétique , microARN/métabolisme , Protéines de liaison à l'ARN/génétique , Vieillissement , Humains , Mâle , Protéine p53 suppresseur de tumeur/métabolismeRÉSUMÉ
Hydrogen sulfide (H2S), a gaseous intracellular signal transducer, participates in multiple physiological and pathological conditions, including reproductive conditions, and disrupts spermatogenesis. The blood-testis barrier (BTB) plays a vital role in spermatogenesis. However, the effect of H2S on the BTB and the underlying mechanism remain unclear. Herein, we examined the effect of H2S and omega-3 polyunsaturated fatty acids (ω-3 PUFAs) on the BTB and testicular functions. ICR male mice were randomly divided into the following groups: control, H2S exposure, and H2S exposure with ω-3 PUFAs intervention. The sperm parameters (sperm concentration and sperm motility) declined in the H2S group and improved in the ω-3 intervention group. BTB integrity was severely disrupted by H2S, and the BTB-related gene levels (ZO-1, Occludin, Claudin 11) decreased; ω-3 supplementation could alleviate BTB disruption by upregulating BTB-related genes, and TM4 Sertoli cells had a similar trend in vitro. p38 MAPK phosphorylation was upregulated in the Na2S treatment group and downregulated after ω-3 cotreatment. These findings suggest that H2S can impair the BTB and that ω-3 PUFAs supplementation can attenuate H2S toxicity in the male reproductive system. Our study elucidated the relationship between a gasotransmitter (H2S) and the BTB and identified the potential therapeutic effect of ω-3 PUFAs.
Sujet(s)
Barrière hématotesticulaire/effets des médicaments et des substances chimiques , Acides gras omega-3/pharmacologie , Sulfures/toxicité , Animaux , Barrière hématotesticulaire/métabolisme , Lignée cellulaire , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Mâle , Souris de lignée ICR , Numération des spermatozoïdes , Mobilité des spermatozoïdes/effets des médicaments et des substances chimiques , Spermatozoïdes/effets des médicaments et des substances chimiques , Testostérone/sang , Protéines de la jonction serrée/génétique , p38 Mitogen-Activated Protein Kinases/métabolismeRÉSUMÉ
HONO measurement was conducted using a wet-chemistry-based method at the Changzhou Environmental Monitoring Center in April 2017. HONO ranged from 0.2-13.9 µg·m-3 with an average of (2.9±2.3) µg·m-3. O3, HCHO, volatile organic compounds, photolysis frequency, and meteorological parameters were simultaneously monitored.·OH concentration was simulated by a Master Chemical Mechanism box model and the daytime maximum·OH concentration ranged from 1.0×106 to 14×106 molecules per cubic centimeter. The formation rates of·OH by photolysis of HONO, O3, HCHO, H2O2, and alkene ozonolysis were calculated as well. The effects of the five sources on atmospheric oxidation capacity were revealed:O3 photolysis (46.4%) > HONO photolysis (41.1%) > alkene ozonolysis (10.9%) > HCHO photolysis (1.5%) > H2O2 photolysis (0.1%). HONO photolysis for OH radical production played a major role in the early morning, before with an increase in O3 concentration, O3 photolysis began to account for most of the·OH production. After 17:00, due to a significant decrease in the intensity of solar radiation, the alkene ozonolysis started playing a major role in the formation of·OH. The photolysis of formaldehyde and hydrogen peroxide played a negligible role in·OH radical production in this study.
RÉSUMÉ
BACKGROUND: Many factors may lead to sperm DNA damage. However, it is little known that the correlations of sperm DNA damage with obesity-associated markers, and reproductive hormones and lipids levels in serum and seminal plasma. METHODS: In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from August 2012 through June 2015. Their obesity-associated markers, semen parameters, sperm acrosomal enzyme activity, seminal plasma biochemical markers, and reproductive hormones and lipids levels in serum and seminal plasma were detected. Sperm DNA fragmentation index (DFI) was determined by sperm chromatin structure assay. The correlations between DFI and each of the above-mentioned variables were analyzed. RESULTS: Spearman correlation analysis showed that sperm DFI was positively related to age and abstinence time (P<0.001). Sperm DFI was also positively related to semen volume and percent of abnormal sperm head (P<0.001), while negatively related to sperm concentration, progressive motility (PR), sperm motility, total normal-progressively motile sperm count (TNPMS), percent of normal sperm morphology (NSM), percent of intact acrosome and acrosomal enzyme activity (P<0.001). Sperm DFI was positively related to seminal plasma zinc level (P<0.001) but unrelated to seminal plasma total α-glucotase, γ-glutamyl transpeptidase (GGT) and fructose levels. There was no any correlation between sperm DFI and obesity-associated markers such as body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC) and waist-to-height ratio (WHtR), and serum lipids levels, but there was positive correlation between sperm DFI and seminal plasma triglyceride (TG) and total cholesterol (TC) levels (P<0.001). Sperm DFI was positively related to serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels and seminal plasma FSH and estradiol (E2) levels (P<0.001), but unrelated to serum and seminal plasma testosterone (T) levels. The multivariate regression analysis for the variables which were significantly correlated with sperm DFI in Spearman correlation analysis showed that age, semen volume, sperm concentration, progressive motility, TNPMS and intact acrosome were independently correlated with sperm DFI. CONCLUSIONS: There are many potential factors associated with sperm DFI, including age, abstinence time, spermatogenesis and maturation, seminal plasma lipids and reproductive hormones levels. However, the potential effects of seminal plasma lipids and reproductive hormones on sperm DNA damage need still to be demonstrated by the studies with scientific design and a large size of samples.
Sujet(s)
Fragmentation de l'ADN , Infertilité masculine , Spermatozoïdes/composition chimique , Acrosome/ultrastructure , Adolescent , Adulte , Facteurs âges , Anthropométrie , Marqueurs biologiques , Chine , Altération de l'ADN , Oestradiol/sang , Hormone folliculostimulante/sang , Humains , Lipides/analyse , Hormone lutéinisante/sang , Mâle , Adulte d'âge moyen , Obésité/physiopathologie , Sperme/composition chimique , Abstinence sexuelle , Numération des spermatozoïdes , Mobilité des spermatozoïdes , Spermatozoïdes/malformations , Triglycéride/analyse , Zinc/analyseRÉSUMÉ
INTRODUCTION: Hyperactivity of the sympathetic nervous system can play an important role in lifelong premature ejaculation (PE). Our previous study found that amyloid precursor protein (APP) levels in seminal plasma of patients with PE were clearly increased. Amyloid-ß (Aß) is derived from APP. Excessive Aß, especially Aß42, can cause neuronal dysfunction. AIM: To determine whether APP and Aß42 are associated with an abnormal penile sympathetic skin response (PSSR). METHODS: From November 2015 to April 2016, 24 patients with lifelong PE (mean age = 29.2 ± 5.3) with self-estimated intravaginal ejaculatory latency time no longer than 2 minutes and 10 control subjects (mean age = 28.0 ± 5.5) were enrolled consecutively from andrology clinics. PSSR was measured in patients with lifelong PE. APP and Aß42 levels in seminal plasma were determined. MAIN OUTCOME MEASURES: PSSR in patients with lifelong PE and APP and Aß42 levels in all subjects. RESULTS: Patients with PE presented 1.5-fold higher levels of APP (P = .004) than control subjects. Seminal plasma protein concentration (C) in the PE group was lower than that in the control group (P = .007). APP divided by C (APP/C) was 2.0-fold higher (P < .001) in the PE group. Aß42 level was not different between the PE and control groups, but Aß42 divided by C (Aß42/C) was significantly higher in the PE group (P < .001). No differences in APP and APP/C were found between patients with PE in the abnormal and normal PSSR groups. The abnormal PSSR group presented significantly higher Aß42 (P = .007) and Aß42/C (P < .001) levels. The latency of PSSR was negatively correlated with Aß42/C (r = -0.436; P = .033). CONCLUSION: These results showed that patients with lifelong PE had higher APP and Aß42 levels in seminal plasma. Abnormal PSSR was related to a higher Aß42 level. Drugs that decrease Aß could be treatment of PE.
Sujet(s)
Éjaculation précoce/physiopathologie , Peau/métabolisme , Système nerveux sympathique/métabolisme , Adulte , Andrologie , Études cas-témoins , Éjaculation/physiologie , Humains , Mâle , Pénis/physiopathologie , Sperme , Jeune adulteRÉSUMÉ
Myriad biological factors have been proposed to explain premature ejaculation (PE). However, data correlating PE with seminal vesicles (SVs) are sparse. The study aimed to evaluate the relationship between the size of SV and PE. The cross-sectional study included 44 outpatients with PE and 44 volunteers without PE, and the size of SV was compared. Self-estimated intravaginal ejaculatory latency time, the Premature Ejaculation Diagnostic Tool (PEDT), the International Index of Erectile Function-15, and the National Institutes of Health-Chronic Prostatitis Symptom Index were used for assessment of symptoms. Compared to the control group, the PE group had significantly higher mean anterior-posterior diameter (APD) of SV (P < 0.001). The optimal mean APD of SV cutoff level was 9.25 mm for PE. In the PE group, PEDT was also higher with a mean APD of SV ≥9.25 mm compared with mean APD of SV <9.25 mm. PEDT was significantly correlated with the mean APD of SV (r = 0.326, P = 0.031). The seminal plasma proteins were compared between six PE and six matched control cases by mass spectrometry and it was shown that 102 proteins were at least 1.5-fold up- or down-regulated. Among them, GGT1, LAMC1, and APP were significantly higher in the PE group. These results indicated that men with a larger mean APD of SV might have a higher PEDT score. Transrectal ultrasound of SV should be considered in the evaluation of patients with premature ejaculation. SV might be a potential target for the treatment of patients with PE and ultrasound change in SV.
Sujet(s)
Éjaculation précoce/imagerie diagnostique , Vésicules séminales/imagerie diagnostique , Adulte , Sujet âgé , Vieillissement , Études cas-témoins , Études transversales , Hormones sexuelles stéroïdiennes/sang , Humains , Lipides/sang , Mâle , Adulte d'âge moyen , Valeurs de référence , Protéines du plasma séminal/analyse , Protéines du plasma séminal/génétique , Facteurs socioéconomiques , Échographie , Échographie-doppler couleur , Ablation par ultrasons focalisés de haute intensité par voie rectale , Jeune adulteRÉSUMÉ
Secretin, a gastrointestinal peptide, has been found to be expressed in mouse endometrial stromal cells (mESCs) during early pregnancy. In order to further investigate the function of secretin during embryo implantation, the expression levels of secretin, secretin receptor, cytosolic phospholipase A2 (cPLA2) and membrane prostaglandin E synthase 1 (mPGEs-1) were detected in the mice uterus from day 4 to 8 of pregnancy by real-time PCR, ELISA and in situ hybridization. mESCs isolated and cultured from day 4 of pregnancy were transfected with secretin expression vectors or treated with H89, a PKA inhibitor. Then the expression levels of cPLA2, mPGEs-1 and cAMP responsive element-binding protein (CREB) were detected by real-time PCR and Western blot. The concentration of prostaglandin E2 (PGE2) in the supernatant was determined by ELISA. The result showed that secretin, cPLA2 and mPGEs-1 mRNA expression increased gradually in implantation sites from day 5 to day 7 of pregnancy with the same tendency. The secretin levels in serum were significantly higher on days 6, 7 and 8 of pregnancy than that on day 5 of pregnancy. The concentration of secretin was significantly higher in implantation sites on days 6, 7 than that in non-implantation site on day 5. Transfection of secretin expression vector promoted cPLA2, p-cPLA2 and mPGEs-1 expressions in mESCs, but not PGE2 level in the supernatant. H89 could effectively inhibit the expression of CREB, p-CREB, p-cPLA2 and cPLA2 induced by secretin. The results showed that the increased secretin expression in mESCs during embryo implantation may promote p-cPLA2, cPLA2 and mPGEs-1 expression, and the promotion may be through PKA signaling pathway.
Sujet(s)
Cellules stromales , Animaux , Technique de Western , Protéine de liaison à l'élément de réponse à l'AMP cyclique , Dinoprostone , Femelle , Souris , Cytosolic phospholipases A2 , Grossesse , Prostaglandin-E synthases , Réaction de polymérisation en chaine en temps réel , Sécrétine , UtérusRÉSUMÉ
OBJECTIVE: This prospective study was designed to investigate the relationship between lipids levels in both serum and seminal plasma and semen parameters. METHODS: 631 subfertile men were enrolled. Their obesity-associated markers were measured, and semen parameters were analyzed. Also, seminal plasma and serum TC, TG, HDL and LDL and serum FFA, FSH, LH, total testosterone (TT), estradiol (E2) and SHBG levels were detected. RESULTS: Seminal plasma and serum TG, TC and LDL levels were positively related to age. Serum TC, TG and LDL were positively related to obesity-associated markers (P < 0.001), while only seminal plasma TG was positively related to them (P < 0.05). For lipids levels in serum and seminal plasma, only TG level had slightly positive correlation between them (r = 0.081, P = 0.042). There was no significant correlation between serum lipids levels and semen parameters. However, seminal plasma TG, TC, LDL and HDL levels were negatively related to one or several semen parameters, including semen volume (SV), sperm concentration (SC), total sperm count (TSC), sperm motility, progressive motility (PR) and total normal-progressively motile sperm counts (TNPMS). Moreover, seminal plasma TG, TC, LDL and HDL levels in patients with oligospermatism, asthenospermia and teratozoospermia were higher than those with normal sperm concentration, motility or morphology. After adjusting age and serum LH, FSH, TT, E2 and SHBG levels, linear regression analysis showed that SV was still significantly correlated with seminal plasma LDL (P = 0.012), both of SC and TSC with seminal plasma HDL (P = 0.028 and 0.002), and both of PR and sperm motility with seminal plasma TC (P = 0.012 and 0.051). CONCLUSION: The abnormal metabolism of lipids in male reproductive system may contribute to male factor infertility.
Sujet(s)
Infertilité masculine/métabolisme , Lipides/analyse , Sperme/composition chimique , Adolescent , Adulte , Vieillissement/métabolisme , Chine , Oestradiol/sang , Acide gras libre/sang , Hormone folliculostimulante/sang , Humains , Infertilité masculine/sang , Infertilité masculine/complications , Lipides/sang , Hormone lutéinisante/sang , Mâle , Adulte d'âge moyen , Obésité/complications , Obésité/métabolisme , Études prospectives , Globuline de liaison aux hormones sexuelles/analyse , Numération des spermatozoïdes , Mobilité des spermatozoïdes , Spermatozoïdes/malformations , Testostérone/sang , Jeune adulteRÉSUMÉ
BAZ1A, a non-catalytic subunit of the chromatin remodeler complexes ACF and CHRAC, is thought to modulate the ATPase's activity of the complexes and participate in gene transcription, DNA damage checkpoint and double-strand break repair. Recently, the essential role of BAZ1A in mouse male fertility has also been reported. BAZ1A contains one C-terminal bromodomain, which specifically recognizes acetylation of lysine. Here, we report the backbone and side chain (1)H, (13)C and (15)N resonance assignment of the mouse BAZ1A-bromodomain, as a basis for further functional studies and structure determination.
Sujet(s)
Protéines chromosomiques nonhistones/composition chimique , Résonance magnétique nucléaire biomoléculaire , Animaux , Souris , Domaines protéiquesRÉSUMÉ
This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3ß-HSD, and 17ß-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3ß-HSD, and 17ß-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 µmol l-1 ) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3ß-HSD, and 17ß-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3ß-HSD, and 17ß-HSD in Leydig cells.
Sujet(s)
Annexine A5/pharmacologie , Antienzymes/pharmacologie , Cellules de Leydig/effets des médicaments et des substances chimiques , Système de signalisation des MAP kinases/effets des médicaments et des substances chimiques , ARN messager/effets des médicaments et des substances chimiques , Testostérone/biosynthèse , 17-Hydroxysteroid dehydrogenases/effets des médicaments et des substances chimiques , 17-Hydroxysteroid dehydrogenases/génétique , 17-Hydroxysteroid dehydrogenases/métabolisme , 3-Hydroxysteroid dehydrogenases/effets des médicaments et des substances chimiques , 3-Hydroxysteroid dehydrogenases/génétique , 3-Hydroxysteroid dehydrogenases/métabolisme , Animaux , Technique de Western , Cholesterol side-chain cleavage enzyme/effets des médicaments et des substances chimiques , Cholesterol side-chain cleavage enzyme/génétique , Cholesterol side-chain cleavage enzyme/métabolisme , Cellules de Leydig/métabolisme , Mâle , Phosphoprotéines/effets des médicaments et des substances chimiques , Phosphoprotéines/génétique , Phosphoprotéines/métabolisme , ARN messager/métabolisme , Rats , RT-PCRRÉSUMÉ
This study investigated the effect of annexin V on the proliferation of primary rat Leydig cells and the potential mechanism. Our results showed that annexin V promoted rat Leydig cell proliferation and cell cycle progression in a dose- and time-dependent manner. Increased level of annexin V also enhanced Ect2 protein expression. However, siRNA knockdown of Ect2 attenuated annexin V-induced proliferation of rat Leydig cells. Taken together, these data suggest that increased level of annexin V induced rat Leydig cell proliferation and cell cycle progression via Ect2. Since RhoA activity was increased following Ect2 activation, we further investigated whether Ect2 was involved in annexin V-induced proliferation via the RhoA/ROCK pathway, and the results showed that annexin V increased RhoA activity too, and this effect was abolished by the knockdown of Ect2. Moreover, inhibition of the RhoA/ROCK pathway by a ROCK inhibitor, Y27632, also attenuated annexin V-induced proliferation and cell cycle progression. We thus conclude that Ect2 is involved in annexin V-induced rat Leydig cell proliferation through the RhoA/ROCK pathway.
Sujet(s)
Tumeur à cellules de Leydig/génétique , Protéines proto-oncogènes/génétique , rho-Associated Kinases/génétique , Protéine G RhoA/génétique , Amides/administration et posologie , Animaux , Annexine A5/administration et posologie , Prolifération cellulaire/effets des médicaments et des substances chimiques , Régulation de l'expression des gènes tumoraux/effets des médicaments et des substances chimiques , Humains , Tumeur à cellules de Leydig/traitement médicamenteux , Tumeur à cellules de Leydig/anatomopathologie , Mâle , Protéines proto-oncogènes/biosynthèse , Pyridines/administration et posologie , Petit ARN interférent , Rats , Transduction du signal/effets des médicaments et des substances chimiques , rho-Associated Kinases/antagonistes et inhibiteurs , rho-Associated Kinases/biosynthèse , Protéine G RhoA/biosynthèseRÉSUMÉ
OBJECTIVE: To investigate the protective effect of epigallocatechin gallate (EGCG) on mouse sperm in vivo. METHODS: A total of 64 six-week-old male Kuming mice were randomly divided into eight groups of equal number to be treated with normal saline (negative control), Cyclophosphamide (CP) at 30 mg/kg (positive control), and CP followed by EGCG (experimental) at 20, 40, and 80 mg/kg, respectively, given every other day for 10 days. At 4 and 5 weeks after treatment, the bilateral testes of the mice were harvested for examination of sperm abnormality. RESULTS: EGCG did not increase the rate of CP-induced sperm abnormality in the mice, but reduced it instead with the prolonged time of treatment. CONCLUSION: EGCG protects mouse sperm in vivo.
Sujet(s)
Catéchine/analogues et dérivés , Spermatozoïdes/effets des médicaments et des substances chimiques , Animaux , Catéchine/pharmacologie , Cyclophosphamide/toxicité , Mâle , Souris , Mutagènes/toxicité , Répartition aléatoire , Facteurs tempsRÉSUMÉ
OBJECTIVE: To study the impact of abdominal obesity on the production of male reproductive endocrine hormones. METHODS: This study included 342 male patients at the andrology clinic, aged 19 -47 years and higher than 160 cm. We measured their waistlines, hiplines and waist-hip ratio, detected the levels of serum estradiol (E2), testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and free testosterone (FT) by chemiluminescence and radioimmunoassay, and analyzed the correlation of the waist-hip ratio with the levels of reproductive endocrine hormones. Abdominal obesity was defined as the waist-hip ratio > 0.9. RESULTS: In the 342 male patients, there were 62 cases of abdominal obesity and 280 cases of the normal somatotype (waist-hip ratio < or = 0.9). The waist-hip ratio was negatively correlated with the T level (r = -0.163, P = 0.003) and the T/LH ratio (r = -0.13, P = 0.02). Both the T level and T/LH ratio were significantly reduced in the abdominal obesity patients ([14.51 +/- 4.53] nmol/L and 2.26 +/- 0.36) as compared with the normal somatotype controls ([17.21 +/- 4.23] nmol/L and 4.61 +/- 0.19) (P < 0.05). CONCLUSION: The waist-hip ratio has a significant negative correlation with the T level and T/LH ratio, and the serum T level is significantly lower in men with abdominal obesity than in those of the normal somatotype.
Sujet(s)
Hormone lutéinisante/sang , Obésité abdominale/sang , Testostérone/sang , Rapport taille-hanches , Adulte , Études cas-témoins , Oestradiol/sang , Hormone folliculostimulante/sang , Humains , Mâle , Adulte d'âge moyen , Somatotypes , Jeune adulteRÉSUMÉ
AIMS: Anti-follicle-stimulating hormone (FSH) autoantibody was found to highly correlate with oligospermia and asthenospermia, but the actual effect of FSH autoantibody on spermatogenesis is still unknown. MAIN METHODS: In this study, 21-day rats were immunized seven times with FSH peptides linked with Keyhole Limpet Hemocyanin (KLH) (experimental group) or KLH (control group) every 2 weeks. Luteinizing hormone (LH) and inhibin B level in the immunized rat sera were measured by enzyme-linked immunosorbent assay (ELISA). Apoptosis of spermatogenic cells in the testis was detected by in situ end labeling method (TUNEL), and the mRNAs of Bax, Bcl-2 and Caspase-3 in testis were detected by fluorescent Quantitative PCR. KEY FINDINGS: Compared with the control, serum inhibin B level was significantly decreased at all time points (34.49%, 23.20%, and 37.00%) (p<0.05). There was no difference in the serum LH level between experimental and control groups. FSH peptide immunization increased the apoptosis of spermatogenic cells in the testis that was associated with an imbalance of Bax and Bcl-2 expression and upregulation of Caspase-3. SIGNIFICANCE: These results suggest that FSH autoantibody could cause the reduction of inhibin B, thereby inducing hypospermatogenesis via augment of spermatogenic cell apoptosis.